Isolation and identification of bacteria from gut of late instar larvae of cocoa pod borer (Conopomorpha Cramerella)

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Abstract

Cocoa Pod Borer (CPS), Conopomorpha cramerella (Lepidoptera: Gracillariidae) is a serious pest of cocoa in South-East Asian countries including Malaysia. It feeds on cocoa placenta and pulp around the beans. These affect the bean development and quality and resulted in reduction of bean yield. CPS enter the cocoa pod by tunneling into thick pericarp layer which consists of complex polysaccharides. This suggests that CPS has the ability to digest the polysaccharide content of cocoa pod. There are possibilities of bacteria colonization in CPS gut which symbiotically involve in digestion of complex pericarp layer. Therefore, this study was conducted to isolate and identify the bacteria from digestive gut of late instar larvae of CPS. The sample of CPB larva was collected from infested cocoa pod which obtained from Agriculture Research
Station Quion Hill, Tawau, Sabah. The gut bacteria were isolated on four (4) different media, Potato Dextrose Agar (PDA), Nutrien Agar (NA), TryptiC Soy Agar (TSA) and Plate Count Agar (PCA). The isolated bacteria were identified by examining colony and cellular morphology, Gram staining and MALDI-TOF Mass Spectrometry analysis. Six (6) types of Gram positive bacteria and a Gram negative bacteria were isolated from the gut of CPB larva. Four (4) species of bacteria were identified using MALDI-TOF Mass Spectrometry analysis. The identified bacteria species are Gram negative Pectobacterium betavasculorum and Gram positive Enterococcus mundtii, Kocuria marina, and Bacillus pumilus. The isolated bacteria might play some important role in digestion and adaptation of CPS on cocoa plant.